An Iodinated DAPI‐Based Reagent for Mass Cytometry. (5th November 2020)

• Record Type: Journal Article
• Authors: Rana, Rahul; Chang, Qing; Bassan, Jay; Chow, Sue; Hedley, David; Nitz, Mark
• Is Part Of: Chembiochem. Volume 22:Number 3(2021)
• Page Start: 532
• Page End: 538
• Publication Date: 2020-11-05
• Subjects: DNA dye -- fluorescent probes -- intercalator -- iodine -- mass cytometry; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.202000369
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Multiparametric single‐cell analysis has seen dramatic improvements with the introduction of mass cytometry (MC) and imaging mass cytometry (IMC ™ ). These technologies expanded the number of biomarkers that can be identified simultaneously by using heavy‐isotope‐tagged antibody reagents. Small‐molecule probes bearing heavy isotopes are emerging as additional useful functional reporters of cellular features. Realizing this, we explored the iodination of DAPI to produce a heavy‐atom‐substituted derivative of the commonly used fluorescent DNA stain. Although exhibiting a drastically reduced fluorescence emission profile, I‐DAPI retains strong binding affinity for DNA. I‐DAPI was used to detect cellular DNA in MC and IMC™ assays with comparable efficiency to known Ir‐containing DNA intercalators. This work suggests repurposing well‐known colorimetric stains through simple reactions could be an effective strategy to develop new, functional MC and IMC™ reagents. Abstract : A heavier DAPI : Synthetic incorporation of iodine into DAPI enabled its repurposing for detecting DNA with high‐dimensional single‐cell technologies: mass cytometry and Imaging Mass Cytometry ™ . Staining of single‐cell suspensions and tissue sections demonstrated reliable identification of nucleated cells and visualization of intracellular DNA.